US7601731B2 - Antimicrobial flush solutions - Google Patents
Antimicrobial flush solutions Download PDFInfo
- Publication number
- US7601731B2 US7601731B2 US10/862,830 US86283004A US7601731B2 US 7601731 B2 US7601731 B2 US 7601731B2 US 86283004 A US86283004 A US 86283004A US 7601731 B2 US7601731 B2 US 7601731B2
- Authority
- US
- United States
- Prior art keywords
- edta
- solution
- minocycline
- ethanol
- antimicrobial
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 230000000845 anti-microbial effect Effects 0.000 title claims abstract description 47
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 271
- 239000004599 antimicrobial Substances 0.000 claims abstract description 58
- 229960004023 minocycline Drugs 0.000 claims description 74
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 23
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 18
- 241000222173 Candida parapsilosis Species 0.000 claims description 16
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims description 12
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- IEDVJHCEMCRBQM-UHFFFAOYSA-N trimethoprim Chemical compound COC1=C(OC)C(OC)=CC(CC=2C(=NC(N)=NC=2)N)=C1 IEDVJHCEMCRBQM-UHFFFAOYSA-N 0.000 claims description 5
- 229960001082 trimethoprim Drugs 0.000 claims description 5
- RJQXTJLFIWVMTO-TYNCELHUSA-N Methicillin Chemical compound COC1=CC=CC(OC)=C1C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 RJQXTJLFIWVMTO-TYNCELHUSA-N 0.000 claims description 3
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- SRVJKTDHMYAMHA-WUXMJOGZSA-N thioacetazone Chemical compound CC(=O)NC1=CC=C(\C=N\NC(N)=S)C=C1 SRVJKTDHMYAMHA-WUXMJOGZSA-N 0.000 description 1
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- 230000002537 thrombolytic effect Effects 0.000 description 1
- 229960000790 thymol Drugs 0.000 description 1
- 229960004659 ticarcillin Drugs 0.000 description 1
- OHKOGUYZJXTSFX-KZFFXBSXSA-N ticarcillin Chemical compound C=1([C@@H](C(O)=O)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)C=CSC=1 OHKOGUYZJXTSFX-KZFFXBSXSA-N 0.000 description 1
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- KVSKGMLNBAPGKH-UHFFFAOYSA-N tribromosalicylanilide Chemical compound OC1=C(Br)C=C(Br)C=C1C(=O)NC1=CC=C(Br)C=C1 KVSKGMLNBAPGKH-UHFFFAOYSA-N 0.000 description 1
- ICUTUKXCWQYESQ-UHFFFAOYSA-N triclocarban Chemical compound C1=CC(Cl)=CC=C1NC(=O)NC1=CC=C(Cl)C(Cl)=C1 ICUTUKXCWQYESQ-UHFFFAOYSA-N 0.000 description 1
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- UJMBCXLDXJUMFB-UHFFFAOYSA-K trisodium;5-oxo-1-(4-sulfonatophenyl)-4-[(4-sulfonatophenyl)diazenyl]-4h-pyrazole-3-carboxylate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)C1=NN(C=2C=CC(=CC=2)S([O-])(=O)=O)C(=O)C1N=NC1=CC=C(S([O-])(=O)=O)C=C1 UJMBCXLDXJUMFB-UHFFFAOYSA-K 0.000 description 1
- WGIWBXUNRXCYRA-UHFFFAOYSA-H trizinc;2-hydroxypropane-1,2,3-tricarboxylate Chemical compound [Zn+2].[Zn+2].[Zn+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O WGIWBXUNRXCYRA-UHFFFAOYSA-H 0.000 description 1
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- 229940126085 β‑Lactamase Inhibitor Drugs 0.000 description 1
Images
Classifications
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/48—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with two nitrogen atoms as the only ring hetero atoms
- A01N43/54—1,3-Diazines; Hydrogenated 1,3-diazines
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N31/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
- A01N31/02—Acyclic compounds
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/16—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using chemical substances
- A61L2/18—Liquid substances or solutions comprising solids or dissolved gases
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L29/00—Materials for catheters, medical tubing, cannulae, or endoscopes or for coating catheters
- A61L29/14—Materials characterised by their function or physical properties, e.g. lubricating compositions
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- A—HUMAN NECESSITIES
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L29/00—Materials for catheters, medical tubing, cannulae, or endoscopes or for coating catheters
- A61L29/14—Materials characterised by their function or physical properties, e.g. lubricating compositions
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- A—HUMAN NECESSITIES
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- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N2300/00—Combinations or mixtures of active ingredients covered by classes A01N27/00 - A01N65/48 with other active or formulation relevant ingredients, e.g. specific carrier materials or surfactants, covered by classes A01N25/00 - A01N65/48
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
Definitions
- the present invention relates generally to the fields of medicine and microbiology. More particularly, it concerns methods of reducing microbial organisms from indwelling medical devices, medical equipment and other surfaces.
- CVCs central venous catheters
- CRBSI catheter-related bloodstream infections
- intralumenal colonization is the major source for the migration of organisms leading to bloodstream infections in long-term silicone catheters (Raad et al., 1993)
- recent guidelines by the CDC and Infectious Diseases Society of America have proposed the use of intralumenal antimicrobial lock solutions for the prevention and treatment of CRBSI (Mermel et al., 2001; Centers for Disease Control and Prevention, 2002).
- Most long-term CVCs are typically flushed with heparin.
- M-EDTA minocycline and EDTA
- M-EDTA has been found to be efficacious in preventing CRBSI, this solution may not be applicable given some of the limitations of the real world of clinical practice.
- the M-EDTA lock solution was required to be exposed to the surface of the indwelling medical device, such as the lumen of catheters, for at least 4 hours.
- M-EDTA requires at least 4 hours of dwell time to eradicate organisms that colonize the lumen of the catheter (see in particular data in U.S. Pat. No. 5,362,754, columns 11 and 12, and Tables 3, 4 and 5 as well as in U.S. Pat. No. 5,688,516, columns 15 and 16, and Tables 3, 4, and 5).
- Providing a four hour exposure time to reduce microbes using the M-EDTA solution is usually not possible in critically ill patients who require continuous infusion therapy, including parenteral nutrition.
- compositions and methods for rapid reduction and/or eradication of microbes from indwelling medical devices without interruption of the use of the device in patients for too long a period.
- antimicrobial compositions there is also a need for better and improved antimicrobial compositions.
- the present invention overcomes these and other limitations in the art and provides compositions that reduce or eradicate microbial agents from surfaces wherein the compositions comprise at least one antimicrobial agent, at least one chelator and/or anticoagulant, and at least one alcohol.
- the present invention also provides methods to rapidly reduce or eradicate microbial agents from surfaces.
- antimicrobial solutions comprising at least one alcohol, at least one antimicrobial agent and at least one chelator and/or anticoagulant.
- Antimicrobial agents that are comprised in the solutions of the present invention include antibacterial agents, antifungal agents, antiviral agents as well as antiseptic agents. These components are present in effective amounts to reduce microbial growth.
- the antimicrobial agent is an antibacterial agent. While any antibacterial agent may be used in the preparation of the instant antimicrobial solutions, some non-limiting exemplary antibacterial agent(s) include those classified as aminoglycosides, beta lactams, quinolones or fluoroquinolones, macrolides, sulfonamides, sulfamethaxozoles, tetracyclines, streptogramins, oxazolidinones (such as linezolid), clindamycins, lincomycins, rifamycins, glycopeptides, polymxins, lipo-peptide antibiotics, as well as pharmacologically acceptable sodium salts, pharmacologically acceptable calcium salts, pharmacologically acceptable potassium salts, lipid formulations, derivatives and/or analogs of the above.
- the aminoglycosides are bactericidal antibiotics that bind to the 30S ribosome and inhibit bacterial protein synthesis. They are typically active against aerobic gram-negative bacilli and staphylococci. Exemplary aminoglycosides that may be used in some specific aspects of the invention include amikacin, kanamycin, gentamicin, tobramycin, or netilmicin.
- Beta lactams are a class of antibacterials that inhibit bacterial cell wall synthesis.
- a majority of the clinically useful beta-lactams belong to either the penicillin group (penam) or cephalosporin (cephem) groups.
- the beta-lactams also include the carbapenems (e.g., imipenem), and monobactams (e.g., aztreonam).
- Inhibitors of beta-lactamase such as clavulanic acid and its derivatives are also included in this category.
- Non-limiting examples of the penicillin group of antibiotics that may be used in the solutions of the present invention include amoxicillin, ampicillin, benzathine penicillin G, carbenicillin, cloxacillin, dicloxacillin, piperacillin, or ticarcillin, etc.
- cephalosporins examples include ceftiofur, ceftiofur sodium, cefazolin, cefaclor, ceftibuten, ceftizoxime, cefoperazone, cefuroxime, cefprozil, ceftazidime, cefotaxime, cefadroxil, cephalexin, cefamandole, cefepime, cefdinir, cefriaxone, cefixime, cefpodoximeproxetil, cephapirin, cefoxitin, cefotetan etc.
- beta lactams include mipenem or meropenem which are extremely active parenteral antibiotics with a spectrum against almost all gram-positive and gram-negative organisms, both aerobic and anaerobic and to which Enterococci, B. fragilis , and P. aeruginosa are particularly susceptible.
- beta lactamase inhibitors include clavulanate, sulbactam, or tazobactam.
- the antibacterial solutions may comprise a combination of at least one beta lactam and at least one beta lactamase inhibitor.
- Macrolide antibiotics are another class of bacteriostatic agents that bind to the 50S subunit of ribosomes and inhibit bacterial protein synthesis. These drugs are active against aerobic and anaerobic gram-positive cocci, with the exception of enterococci, and against gram-negative anaerobes. Exemplary macrolides include erythromycin, azithromycin, clarithromycin.
- Quinolones and fluoroquinolones typically function by their ability to inhibit the activity of DNA gyrase. Examples include nalidixic acid, cinoxacin, trovafloxacin, ofloxacin, levofloxacin, grepafloxacin, trovafloxacin, sparfloxacin, norfloxacin, ciprofloxacin, moxifloxacin and gatifloxacin.
- Sulphonamides are synthetic bacteriostatic antibiotics with a wide spectrum against most gram-positive and many gram-negative organisms. These drugs inhibit multiplication of bacteria by acting as competitive inhibitors of p-aminobenzoic acid in the folic acid metabolism cycle. Examples include mafenide, sulfisoxazole, sulfamethoxazole, and sulfadiazine.
- the tetracycline group of antibiotics include tetracycline derivatives such as tigecycline which is an investigational new drug (IND), minocycline, doxycycline or demeclocycline and analogs such as anhydrotetracycline, chlorotetracycline, or epioxytetracycline.
- IND investigational new drug
- minocycline doxycycline
- demeclocycline analogs such as anhydrotetracycline, chlorotetracycline, or epioxytetracycline.
- EDTA is unique in effectively preventing and dissolving polysaccharide-rich microbial glycocalyx (U.S. Pat. No. 5,362,754).
- streptogramin class of antibacterial agents is exemplified by quinupristin, dalfopristin or the combination of two streptogramins.
- Drugs of the rifamycin class typically inhibit DNA-dependent RNA polymerase, leading to suppression of RNA synthesis and have a very broad spectrum of activity against most gram-positive and gram-negative bacteria including Pseudomonas aeruginosa and Mycobacterium species.
- An exemplary rifamycin is rifampicin.
- antibacterial drugs are glycopeptides such as vancomycin, teicoplanin and derivatives thereof.
- antibacterial drugs are the polymyxins which are exemplified by colistin.
- metronidazole is active only against protozoa, such as Giardia lamblia, Entamoeba histolytica and Trichomonas vaginalis , and strictly anaerobic bacteria.
- Spectinomycin is a bacteriostatic antibiotic that binds to the 30S subunit of the ribosome, thus inhibiting bacterial protein synthesis and nitrofurantoin is used orally for the treatment or prophylaxis of UTI as it is active against Escherichia coli, Klebsiella - Enterobacter species, staphylococci, and enterococci.
- the antimicrobial agent is an antifungal agent.
- Some exemplary classes of antifungal agents include imidazoles or triazoles such as clotrimazole, miconazole, ketoconazole, econazole, butoconazole, omoconazole, oxiconazole, terconazole, itraconazole, fluconazole, voriconazole (UK 109,496), posaconazole, ravuconazole or flutrimazole; the polyene antifungals such as amphotericin B, liposomal amphoterecin B, natamycin, nystatin and nystatin lipid formualtions; the cell wall active cyclic lipopeptide antifungals, including the echinocandins such as caspofungin, micafungin, anidulfungin, cilofungin; LY121019; LY303366; the allylamine group of antifungals such as terbin
- antifungal agents include naftifine, tolnaftate, mediocidin, candicidin, trichomycin, hamycin, aurefungin, ascosin, ayfattin, azacolutin, trichomycin, levorin, heptamycin, candimycin, griseofulvin, BF-796, MTCH 24, BTG-137586, pradimicins (MNS 18184), benanomicin; ambisome; nikkomycin Z; flucytosine, or perimycin.
- the antimicrobial agent is an antiviral agent.
- antiviral agents include cidofovir, amantadine, rimantadine, acyclovir, gancyclovir, pencyclovir, famciclovir, foscamet, ribavirin, or valcyclovir.
- the antimicrobial agent is an innate immune peptide or proteins.
- Some exemplary classes of innate peptides or proteins are transferrins, lactoferrins, defensins, phospholipases, lysozyme, cathelicidins, serprocidins, bacteriocidal permeability increasing proteins, amphipathic alpha helical peptides, and other synthetic antimicrobial proteins.
- the antimicrobial agent is an antiseptic agent.
- antiseptic agents include a taurinamide derivative, a phenol, a quaternary ammonium surfactant, a chlorine-containing agent, a quinaldinium, a lactone, a dye, a thiosemicarbazone, a quinone, a carbamate, urea, salicylamide, carbanilide, a guanide, an amidine, an imidazoline biocide, acetic acid, benzoic acid, sorbic acid, propionic acid, boric acid, dehydroacetic acid, sulfurous acid, vanillic acid, esters of p-hydroxybenzoic acid, isopropanol, propylene glycol, benzyl alcohol, chlorobutanol, phenylethyl alcohol, 2-bromo-2-nitropropan-1,3-diol, formaldehyde, glutaralde
- the antiseptic agent is as set forth in the specification of U.S. Provisional Application Ser. No. 60/261,447, U.S. Provisional Application Ser. No. 60/316,165, and U.S. Non-Provisional patent application Ser. No. 10/044,842, incorporated herein by reference in their entirety.
- the antiseptic agent comprises a basic reagent and a dye.
- the basic reagent may be a guanidium compound, a biguanide, a bipyridine, a phenoxide antiseptic, an alkyl oxide, an aryl oxide, a thiol, a halide, an aliphatic amine, or an aromatic amine.
- the basic reagent is a guanidium compound.
- Non-limiting examples of guanidium compounds include chlorhexidine, alexidine, hexamidine.
- the basic reagent is a bipyridine.
- One example of a bipyridine is octenidine.
- the basic reagent is a phenoxide antiseptic.
- the dye may be a triarylmethane dye, a monoazo dye, a diazo dye, an indigoid dye, a xanthene dye, an anthraquinone dye, a quinoline dye, an FD&C dye.
- triarylmethane dye include gentian violet, crystal violet, ethyl violet, or brilliant green.
- Other non-limiting examples of FD&C dye include Blue No. 1 or Green No. 3.
- One non-limiting example of diazo dyes is D&C Red No. 17.
- An example of an indigoid dye is FD&C Blue No. 2.
- An examples of a xanthene dye is FD&C Red No. 3; of an anthraquinone dye is D&C Green No. 6; and of an quinoline dye is D&C Yellow No. 1.
- antiseptic agents that may be used to prepare the amntimicrobial solutions of the invention are gendine, genlenol, genlosan, or genfoctol.
- antimicrobial agents including one or more antibacterial agent, and/or one or more antifungal agent, and/or one or more antiviral agent, and/or one or more antiseptic agent, and/or combinations thereof.
- chelator agents are contemplated as useful in preparing the antimicrobial solutions of the invention. This includes chelators such as EDTA free acid, EDTA 2Na, EDTA 3Na, EDTA 4Na, EDTA 2K, EDTA 2Li, EDTA 2NH 4 , EDTA 3K, Ba(II)-EDTA, Ca(II)-EDTA, Co(II)-EDTACu(II)-EDTA, Dy(III)-EDTA, Eu(III)-EDTA, Fe(III)-EDTA, In(III-EDTA, La(III)-EDTA, CyDTA, DHEG, diethylenetriamine penta acetic acid (DTPA), DTPA-OH, EDDA, EDDP, EDDPO, EDTA-OH, EDTPO, EGTA, HBED, HDTA, HIDA, IDA, Methyl-EDTA, NTA, NTP, NTPO, O-Bistren, TT
- at least one anticoagulant such as heparin, hirudin, EGTA, EDTA, urokinase, streptokinase, hydrogen peroxide etc.
- Alcohols are contemplated as useful in the preparation of the instant antimicrobial solution, and include any antimicrobially active alcohol.
- Non-limiting examples of alcohols include ethanol, methanol, isopropanol, propylene glycol, benzyl alcohol, chlorobutanol, phenylethyl alcohol, and the like.
- the concentration of the alcohol is preferably in the range of 5%-80% (v/v), more preferably in the range of 10% to 50%, more preferably in the range of 15% to 40%, more preferably in the range of 20% to 30%, with the most preferable being about 25%.
- the more preferred concentration of alcohol will include 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or 80% (v/v) of the alcohol in the preparation of the instant antimicrobial solutions.
- the solutions of the instant invention can comprise various combinations of at least one alcohol, at least one antimicrobial agent, and at least one chelator/anticoagulant.
- the solution of the invention comprises at least one alcohol, at least one tetracycline and at least one chelator/anticoagulant.
- such an antimicrobial solution comprises ethanol, at least one tetracycline and EDTA or heparin.
- such a solution comprises ethanol, minocycline and EDTA or heparin.
- the concentration of minocycline is 0.001 mg/ml to 100 mg/ml.
- the concentration of minocycline is about 3 mg/ml.
- the concentration of EDTA is in the range of 10-100 mg/ml. In one embodiment of this aspect, the concentration of EDTA is about 30 mg/ml.
- the invention also provides methods for reducing microbial organisms from a surface comprising: a) obtaining an antimicrobial solution of the invention as set forth above; and b) contacting the surface with the antimicrobial solution, whereby said contacting reduces microbial organisms from the surface.
- the contacting is performed for 4 hours or less. In other embodiments of the method, the contacting is performed for 2 hours or less, for 1 hour or less, for 30 minutes or less, or for 15 minutes or less.
- the method further comprises eradicating microbes from the surface wherein the contacting is performed for about 15 minutes or more.
- the methods of the invention can be used to reduce microbial agents from the surface of a medical device such as a catheter, an endotracheal tube, a nephrostomy tube, a biliary stent, an orthopedic device, a prosthetic valve, a medical implant, dental devices or dental implants, cardiac assist devices, vascular grafts, tracheostomy, ventriclulostomy devices, or intrathecal devices.
- a medical device such as a catheter, an endotracheal tube, a nephrostomy tube, a biliary stent, an orthopedic device, a prosthetic valve, a medical implant, dental devices or dental implants, cardiac assist devices, vascular grafts, tracheostomy, ventriclulostomy devices, or intrathecal devices.
- the catheter is an indwelling catheter such as a central venous catheter, a peripheral intravenous catheter, an arterial catheter, a Swan-Ganz catheter, a hemodialysis catheter, an urinary catheter, a peritoneal catheter, an umbilical catheter, a percutaneous nontunneled silicone catheter, a cuffed tunneled central venous catheter or a subcutaneous central venous port.
- a central venous catheter such as a central venous catheter, a peripheral intravenous catheter, an arterial catheter, a Swan-Ganz catheter, a hemodialysis catheter, an urinary catheter, a peritoneal catheter, an umbilical catheter, a percutaneous nontunneled silicone catheter, a cuffed tunneled central venous catheter or a subcutaneous central venous port.
- the methods of the invention are useful in reducing microbial agents from a surface such as an organic surface or an inorganic surface.
- a surface such as an organic surface or an inorganic surface.
- An organic surface is exemplified by skin, surgical sutures, mucosal membrane surface, or an epithelial surface.
- An inorganic surface may be the surface of a pipe or pipeline, a floor, a table-top, a counter-top, hospital equipment, or a wheel chair, etc.
- a pipe is an oil pipeline, a water pipeline, an ice machine pipe, or a beverage dispensing pipe.
- antimicrobial solutions of the present invention will find particular usefulness as antimicrobial mouthwash solutions.
- Such mouthwash solutions are contemplated to be useful both in conjunction with dental procedures and oral sterilization as well as in general dental and oral hygiene applications.
- Antimicrobial mouthwash is becoming extremely important in the prevention of oral cavity infections as well as aspiration pneumonia. Microbial organisms in the mouth particularly around the teeth, embed themselves in biofilm and the pathogenesis of infection and colonization is similar to that seen in, for example, vascular catheters.
- one will preferably apply the triple combinations of the present invention, that will include an antimicrobial (possibly antiseptic) with EDTA and low concentration alcohol as a mouthwash or mouth flush solution.
- the invention also provides a kit for disinfecting a surface to reduce microorganisms thereon, wherein the kit comprises components including at least one antimicrobial agent, at least one anticoagulant/chelator, and at least one alcohol, contained in a suitable container.
- the components may be combined in a single container, or powdered components may be lyophilized, combined and separately compartmentalized, or all of the components may be placed in separate containers.
- only the antimicrobial agent(s) is included as a dried powder.
- the kit may optionally include a second carrier solution for reconstituting the lyophilized antibiotic agent(s).
- the kit will include a unit dose of a pharmacologically effective amount of minocycline and EDTA (or heparin), either provided separately as a lyophilized or powdered dose or already mixed in an ethanol solution.
- the unit dose contains at least about 9 mg of minocycline and at least about 90 mg of EDTA.
- Such a kit may further comprise a preselected amount of an ethanol solution such that when the ethanol solution is mixed with the lyophilized unit dose, the concentration of minocycline is 3 mg/ml and the concentration of EDTA is 30 mg/ml.
- Kits in accordance with the present invention may be used to reduce/eliminate microbes on the surface of a medical device, a pipe or pipeline, a floor, a table-top, a counter-top, hospital equipment, or a wheel chair. It is also contemplated that the kits of the invention will further comprise a means for introducing the kit components into the medical device, the pipe or surface.
- a syringe or vial comprising a lyophilized unit dose of a pharmacologically effective amount of one or more of the three components of the flush solutions of the present invention.
- a syringe may comprise minocycline and EDTA (or heparin) mixed in an ethanol solution.
- the unit dose contains at least about 9 mg of minocycline and at least about 90 mg of EDTA.
- Such a syringe or vial may further comprises a preselected amount of an ethanol solution such that when the ethanol solution is mixed with the lyophilized unit dose, the desired concentration of the particular agent is obtained, such as about 3 mg/ml in the case of minocycline and about 30 mg/ml. in the case of EDTA.
- a locking solution for filling and/or flushing a medical indwelling device such as, but not limited to, an implanted catheter
- the locking solution may comprise at least one antimicrobial agent, at least one chelator and/or anticoagulant, and at least one alcohol.
- an “antimicrobial agent” is defined herein as an agent that has antibiotic properties against bacteria, fungi, viruses and other pathogens and includes antibacterial agents, antifungal agents, antiviral agents and antiseptic agents.
- antifungal agent is defined as a compound having either a fungicidal or fungistatic effect upon fungi contacted by the compound.
- fungicidal is defined to mean having a destructive killing action upon fungi.
- fungistatic is defined to mean having an inhibiting action upon the growth of fungi.
- antibacterial agent is defined as a compound having either a bactericidal or bacteriostatic effect upon bacteria contacted by the compound.
- bactericidal is defined to mean having a destructive killing action upon bacteria.
- bacteriostatic is defined to mean having an inhibiting action upon the growth of bacteria.
- antiviral agent is defined as a compound that can either kill viral agents or one that stops the replication of viruses upon contact by the compound.
- the phrase “effective amount” or “therapeutically effective amount” is defined as a dosage sufficient to induce a microbicidal or microbistatic effect upon the microbes contacted by the composition on a surface.
- a chelator denotes one or more chelators.
- the term “chelator” is defined as a molecule comprising nonmetal atoms, two or more of which atoms are capable of linking or binding with a metal ion to form a heterocyclic ring including the metal ion.
- contact As used herein the terms “contact”, “contacted”, and “contacting”, or “exposed” and “exposure” are used to describe the process by which any of the compositions disclosed in the present invention, comes in direct juxtaposition with the surface of a medical device or any other surface from which microbial growth is to be reduced or eradicated.
- a” or “an” may mean one or more.
- the words “a” or “an” when used in conjunction with the word “comprising”, the words “a” or “an” may mean one or more than one.
- another may mean at least a second or more.
- FIG. 1 Ethanol in combination with M-EDTA as a flush solution used for 15 minutes or 24 hours, as indicated, tested against MRSA in biofilm.
- Microorganisms that attach themselves to inert surfaces such as medical devices including, vascular catheters, endotracheal tubes, Foley catheters, biliary stents, nephrostomy tubes, prosthetic valves, ventriculostomy or epidural catheters, or fluid pipelines, such as oil pipelines or water pipelines, produce a layer made of exopolysaccharide called microbial biofilm. These organisms embed themselves in this layer. This biofilm layer ultimately becomes the protective environment that shields these organisms on the inert surface from the antimicrobial activity of various antibiotics or antiseptics.
- medical devices including, vascular catheters, endotracheal tubes, Foley catheters, biliary stents, nephrostomy tubes, prosthetic valves, ventriculostomy or epidural catheters, or fluid pipelines, such as oil pipelines or water pipelines.
- the present invention allows rapid reduction and/or eradication of microorganisms embedded in a biofilm in a time as short as about 15 minutes of exposure to combinations of at least one antimicrobial and at least one chelator/anticoagulant, if this combination is prepared in an alcohol.
- This is exemplified in one embodiment by minocycline-EDTA in an ethanol solution, which is described in detail in the application.
- minocycline-EDTA in an ethanol solution, which is described in detail in the application.
- any antimicrobial agent, any chelator/anticoagulant and any alcohol may be any antimicrobial agent, any chelator/anticoagulant and any alcohol.
- the present invention provides antimicrobial solutions comprising one or more antimicrobial agents, one or more chelator/anticoagulant, and an alcohol solution.
- the present invention also provides methods for the rapid reduction or eradication of microorganisms embedded in a biofilm on a surface comprising contacting or exposing the surface to a flush solution of the invention.
- the invention provides methods for reducing or eradicating microbes from the surfaces of medical devices, including indwelling medial devices, as well as other surfaces, pipelines and the like.
- compositions and the methods of the present invention have an unexpected and surprising efficacy not provided by compositions that comprise only alcohol solutions, or compositions that comprise combinations of antimicrobials with chelators/anticoagulants.
- the combination of the antimicrobial agent minocycline with the chelator/anticoagulant EDTA requires about 4 hours of exposure or dwell time to reduce microbes from the surface of a medical device.
- a 25% ethanol solution alone suppresses colonizing organisms embedded in biofilm, but does not eradicate them.
- one exemplary composition of the present invention comprising minocycline, EDTA and 25% ethanol provides rapid reduction and/or eradication of the microbial organisms within 15 minutes of exposure and also prevents the re-growth of the microbes.
- One of the applications of the antimicrobial flush solutions of the invention is to reduce or eradicate microbes from the surfaces of medical devices especially indwelling medical devices such as catheters, endotracheal tubes, nephrostomy tubea, biliary stents, orthopedic devices, prosthetic devices, and medical implants.
- indwelling medical devices such as catheters, endotracheal tubes, nephrostomy tubea, biliary stents, orthopedic devices, prosthetic devices, and medical implants.
- venous catheters There are at least 5 million central venous catheters inserted annually in the United States, 1.5 million of which are long-term catheters that remain in place for an average of 100 days, and at least 3.5 million short-term catheters that remain for an average of 7 days. All of these venous catheters are flushed with heparin on a daily basis. It is estimated that at least 150-175 million catheter flushes occur annually in the United States alone. Heparin has good anticoagulant activity and, hence, prevents thrombotic occlusions. However, heparin has no antimicrobial activity and, in fact, given the alkaline media that heparin creates, it has been shown to be a promoter of microbial colonization of catheter surfaces.
- indwelling vascular catheters Irrespective of whether heparin is used, almost 90%-100% of indwelling vascular catheters end up being colonized with organisms embedded in biofilm on the surface of these devices, particularly at the lumenal surface. Hence, the most serious and frequent complication of vascular catheters is infection, whereby as fluid is flushed through the lumen of the catheter, microorganisms migrate into the bloodstream and cause catheter-related bloodstream infections. Indwelling central venous catheters are associated with around 5%-8% frequency of catheter-related bloodstream infection, which in turn is associated with an attributable mortality of 25% in critically ill patients. Such an event is also associated with high morbidity and a cost per episode of an average of $30,000.
- EDTA is a well-known chelator of iron and calcium, as well as an active anticoagulant used in blood collection tubes. EDTA has been shown to have equivalent anticoagulant activity to heparin. In addition, EDTA has antibiofilm activity and enhances the antimicrobial activity of other antimicrobial agents, such as minocycline. However, for a combination of an antmicrobial with a chelator (such as minocycline-EDTA) to eradicate organisms embedded in biofilm, contacting the surface for at least 4-hour with this combination is required. This is demonstrated in U.S. Pat. No. 5,362,754 (see especially data in Tables 3, 4 and 5) and in U.S. Pat. No.
- the present invention thus, provides that indwelling medical devices such as catheters be flushed with this antimicrobial and chelator/anticoagulant in an alcohol-based solution.
- This will provide chelation/anticoagulation through the chelator (such as EDTA).
- the combination of antibiotic/chelator with an alcohol results in broad-spectrum reduction or eradication of microbial organisms embedded in biofilm.
- the alcohol further increases the efficacy of the combination.
- Angioplasty devices, heart valves and cardiac pacemakers also are included within the present invention.
- Catheters such as urinary, venous, arterial, and peritoneal catheters may be treated with the flush solutions of the invention.
- tracheotomy devices, shunts, surgical sutures, and other medical devices or prosthesis can be treated.
- the medical devices which are amenable to coatings of the compositions of the invention generally have surfaces composed of thermoplastic or polymeric materials such as polyethylene, Dacron, nylon, polyesters, polytetrafluoroethylene, polyurethane, latex, silicone elastomers and the like.
- Devices with metallic surfaces are also amenable to coatings with the antibiotic combinations. Such devices are exemplified by bone and joint prosthesis. It is also contemplated that the solutions of the invention will be used to disinfect organic surfaces such as skin as well as mucosal surfaces.
- An antimicrobial locking solution of the present invention may comprise at least one alcohol, at least one antimicrobial agent and at least one chelator and/or anticoagulant.
- Various antimicrobial substances as disclosed herein and that are well known to one of ordinary skill in the art may be combined with the locking solution in order to inhibit infection.
- the antimicrobial locking solution of the present invention may be use for filling or flushing a medical device such as an indwelling device such as an implanted catheter. Other medical devices that are contemplated for use in the present invention are disclosed herein.
- the flush solutions of the present invention are also useful in the eradication of the surfaces of other surfaces that microbes can grow on such as pipes, pipelines etc.
- Fluid pipelines such as oil and water pipelines
- lumenal biofilm that is produced by microorganisms that colonize the internal surface of these pipelines.
- these pipelines are flushed with antimicrobial agents.
- antimicrobial and antiseptic agents have little activity against organisms embedded in biofilm. Tons of antibiotics, such as gentamicin, are often used to flush the lumen of oil pipelines, to no avail.
- the present invention provides new and effective compositions and methods for the eradication of organisms, as well as biofilm embedding the lumen of pipelines (oil, water), as well as other devices, such as ice machines.
- These pipelines or machines can be flushed or rinsed with the compositions of the invention that comprise at least one antimicrobial agent and at least one chelator or anticoagulant prepared in a base solution of ethanol. Flushing the pipelines, machines or tubes with the compositions of the invention provide rapid reduction and/or eradication of the biofilm and the organisms in biofilm thereby preventing any obstruction or contamination of the water, oil or the ice machines in certain environmental settings.
- compositions are contemplated to have one or more antimicrobial agents.
- Antimicrobial agents are defined herein as antibacterial agents, antifungal agents, antiviral agents and/or antiseptic agents.
- antimicrobial agent some exemplary classes and examples of antibacterial agents, antifungal agents, antiviral agents as well as antiseptic agents are described above in the section entitled “summary of invention.” Of course one of skill in the art will appreciate that any combination as well as agents from the different types and classes of the antimicrobial agents can be combined to prepare the solutions of the invention.
- Some non-limiting exemplary bacterial and fungal microbes that can be reduced or eradicated by the compositions and methods of the invention include Staphyloccous species such as Staphylococcus epidermidis, Staphylococcus aureus; Aspergllus species, such as Aspergillus flavus, Aspergillus terreus; Fusarium oxysporum, Candida species, such as Candida krusei, Candida parapsilosis, Candida tropicalis, Candida albicans and Candida glabrata .
- viruses can also be eradicated.
- a chelate is the type of coordination compound in which a central metal ion is attached by coordinate links to two or more nonmetal atoms in the same molecule. Heterocyclic rings are thus formed during chelation, with the metal atom as part of the ring.
- the molecule comprising the nonmetal linking atoms is termed a chelator.
- Chelators are used in various chemical applications, for example as titrating agents or as metal ion scavengers. Chelators can be used to remove ions from participation in biological reactions. For example, the well-known chelator ethylenediamine-N,N,N′,N′,-tetraacetic acid (EDTA) acts as an anticoagulant because it is capable of scavenging calcium ions from the blood.
- EDTA ethylenediamine-N,N,N′,N′,-tetraacetic acid
- chelators have significant growth inhibitory effect against several microbes. It is known that iron and other trace metals are essential in the life cycle of microorganisms such as fungi and bacteria. Without these trace metals, microbes are unable to grow and reproduce. Although iron is abundant in nature, its availability for microbial assimilation is limited owing to the insolubility of ferric ions at neutral or alkaline pH. As a consequence, many microbes have evolved their own specialized trace metal-scavenging molecules, called siderophores, which bind with trace metals and make them available for uptake by the microbes.
- the chelators used in conjunction with the present invention provide an inhibitory effect upon microbial pathogens by competing with the siderophores for any available trace metal ions.
- the chelators present in the pharmaceutical preparations of the present invention “steal” the metal ions essential for microbial growth, effectively causing the microbe to “starve to death.”
- the additional antibiotic agents and the ethanol of the compositions of the present invention then come in and attack the weakened microbe, thereby destroying them or inhibiting their growth.
- Table 1 below provides a representative list of chelators useful in conjunction with the present invention. However, the list provided in Table 1 is not meant to be exhaustive.
- Preferred chelators are those which bind trace metal ions with a binding constant ranging from 10 1 to 10 100 . More preferred chelators are those which bind trace metal ions with a binding constant ranging from 10 10 to 10 80 ; and most preferred chelators are those which bind trace metal ions with a binding constant ranging from 10 15 to 10 60 .
- preferred chelators are those which have been shown to have an inhibitory effect upon target microbial pathogens, for example the disodium salt of EDTA.
- anticoagulants such as EGTA, EDTA, heparin, urokinase, streptokinase, low molecular weight heparin, enoxaparin, sodium coumarin, indanedione, anisindione, warfarin, protamine sulfate, anti-thrombin III, nitrilotriacetic acid, potassium sodium tartrate, potassium hydrogen D-tartrate, L-tartaric acid dipotassium salt, L-tartaric acid disodium salt, L-tartaric acid monosodium salt, tris(carboxymethyl)amine, warfarin, acetylsalicylic acid, ibuprofen, indomethacin, prostaglandins, sulfinpyrazone, streptokinase, urokinase, tissue plasminogen activator, coumarin, protamine s
- anticoagulants such as EGTA, EDTA, heparin,
- the flush solutions of the instant invention are contemplated to comprise an alcohol, such as an antiseptic or disinfectant alcohol.
- exemplary alcohols include ethanol, methanol, isopropanol, benzyl alcohol, chlorobutanol, phenylethyl alcohol, 2-bromo-2-nitropropan-1,3-diol, and the like.
- the present invention contemplates any effective concentration of alcohol, but will typically employ a final alcohol concentration in the range of 5%-80% (v/v), more preferably in the range of 10% to 50%, more preferably in the range of 15% to 40%, more preferably in the range of 20% to 30%, with the most preferable being about 25%.
- the more preferred concentration of alcohol will include 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or 80% (v/v) of the alcohol in the preparation of the instant antimicrobial solutions.
- Alcohols such as ethanol are long known to have disinfectant properties.
- EP1245247 and U.S. Pat. No. 6,350,251 it is reported that the combination of ethanol with EDTA provides a biocidal lock for indwelling medical devices.
- a combination of ethanol with EDTA is less effective in killing microbes than ethanol alone (Sherertz et al., 2002).
- the art is in a flux about the exact role of the combination of ethanol with EDTA.
- the present inventor has shown that ethanol alone, while requiring only a relatively short duration of contact, is only partially effective in killing or controlling microbes on the surface of an indwelling medical device or other surface.
- a combination of an antimicrobial agent and a chelator such as EDTA may be effective, yet it requires a somewhat longer duration of contact (e.g., sometimes on the order of 4 hours).
- the triple combination of an alcohol, an antimicrobial and a chelator/anticoagulant provides unexpectedly effective anti-microbial properties in a very short duration and in addition to eradicating microbes rapidly from a surface they also preventing re-growth of the microbial pathogen on the surface.
- An additional advantage for the triple combination is that it is effective at eradicating a broader range of microbial organisms (bacteria and fungi), even at the shorter durations of contact with the treated surface.
- any additional pharmacologically active ingredients or sterilization agents may be comprised in the solutions of the present invention or may be used separately for flushing or treating the devices of the present invention to further reduce or eliminate pathogenic microbes and viruses.
- Typical pharmacologically active ingredients include antifibrin agents, anti-thrombotic agents, and anti-inflammatory agents.
- Anti-inflammatory agents include steroids, and nonsteroidal anti-inflammatory agents, and salicylates.
- Anti-thrombotic drugs including acetylsalicylic acid, dipyridamole, heparin, ibuprofen, indomethacin, prostaglandins, sulfinpyrazone, warfarin, thrombolytic enzymes such as streptokinase, urokinase, or plasminogen activator.
- Complexing agents such as ammonium-1-pyrrolidine dithiocarbanate may also be used.
- the above examples are not meant to be limiting.
- kits will optionally include an instruction sheet insert to identify how the kit is to be used.
- kits described in this section are exemplified by a solution comprising minocycline as the antibiotic, EDTA as the chelator/anticoagulant, and ethanol.
- minocycline as the antibiotic
- EDTA as the chelator/anticoagulant
- ethanol as the chelator/anticoagulant
- any other combination of one or more antibiotic, one or more chelator/anticoagulant, and ethanol as described in the present disclosure may be packaged in a similar manner.
- the kit may comprise of one or two or three or more compartments.
- the components of the kit may be provided in separate compartments or in the same compartment.
- the components of the kit may be provided separately or mixed.
- the mixed components may contain two or more agents such as an antibiotic, a chelator/anticoagulant, or ethanol, or additional component.
- One of the packaging options below maintain the ingredients, for example, the antibiotic, such as minocycline, and the chelating agent/anticoagulant, such as EDTA, in an uncombined form. These components are to be combined shortly before use. These packaging options are contemplated to be part of a 2-compartment or three-compartment container system to provide a total volume of about 3 ml of the ready to use preparation. Any compartmentalized container system may be used to package the compositions of the present invention. An exemplary container system is available from Becton Dickinson.
- Option 1 A 3-Compartment system comprising two dry components such as 3-9 mg minocycline (dry), 10-100 mg EDTA (powdered) and one wet component comprising 3 ml diluent (alcohol alone or diluted in saline or distilled water).
- dry components such as 3-9 mg minocycline (dry), 10-100 mg EDTA (powdered) and one wet component comprising 3 ml diluent (alcohol alone or diluted in saline or distilled water).
- the dry components, minocycline and EDTA will be allowed to mix with the diluent.
- Final concentration of the mixture should be about 3 mg/ml minocycline and 30 mg/ml EDTA.
- Option 2 A 2-Compartment system antibiotic and chelator/anticoagulant (one wet, one dry) comprising for example 3-9 mg/ml minocycline and 10-100 mg EDTA.
- the dry EDTA powder will be combined with the minocycline in solution.
- the minocycline may be suspended in either saline, distilled water, alcohol solution or other physiologically acceptable diluent.
- the minocycline may be in a dry powdered form, and the EDTA in solution.
- a wet/wet® dual chamber container system, available from Becton-Dickinson, may be used in these applications.
- Option 3 A 2 compartment system comprising both wet compartments comprising antimicrobial agent(s) and chelator/anticoagulant comprising in one example 10-100 EDTA Solution and 3-9 mg/ml Minocycline Solution where the solution comprises alcohol.
- the EDTA solution When ready for use, the EDTA solution will be combined with minocycline solution. Once combined, the solution will have a concentration of 3 mg/ml minocycline and 30 mg/ml EDTA.
- a wet/wet® dual chamber container system available from Becton-Dickinson, may be used in these applications.
- the dry EDTA and dry minocycline may be suspended in a solution of an alcohol made in either saline, distilled water, or other physiologically acceptable diluent.
- a liquid/dry® dual container system, from Becton-Dickinson, may be used. When ready for use, the dry minocycline powder will be allowed to combine with the EDTA solution.
- the EDTA can be suspended in either saline or distilled water, or alcohol solution, or other physiologically acceptable diluent.
- kits may include a container means comprising a volume of diluent, comprising an alcohol optionally diluted if required in a solution such as saline or sterile water, a second (or more) container means comprising one or more antimicrobial or biocide, a third (or more) container means comprising one or more chelating/anticoagulant agent.
- a container means comprising a volume of diluent, comprising an alcohol optionally diluted if required in a solution such as saline or sterile water
- a second (or more) container means comprising one or more antimicrobial or biocide
- a third (or more) container means comprising one or more chelating/anticoagulant agent.
- the dry components may optionally be mixed in one compartment.
- the addition of the diluent would then be performed immediately prior to use.
- the container means of the kits will generally include at least one vial, test tube, flask, bottle, syringe or other container means, into which the antimicrobial/chelator/anticoagulant/alcohol may be placed, and preferably, suitably aliquoted. Where a second or third antibiotic agent, other chelator, alcohol, or additional component is provided, the kit will also generally contain a second, third or other additional container into which this component may be placed.
- the kits of the present invention will also typically include a means for containing the alcohol, antimicrobial agent, chelator/anticoagulant, and any other reagent containers in close confinement for commercial sale. Such containers may include injection or blow-molded plastic, or glass containers into which the desired vials are retained.
- the in vitro model utilized a modified Robbins device (MRD) to study the colonization of catheter segments with organisms embedded in biofilm.
- MRD modified Robbins device
- the modified Robbins device has been previously described (Nickel et al., 1991; Evans et al., 1987, see also U.S. Pat. No. 5,362,754) and is constructed from an acrylic block, 42 cm long with a lumen of 2 ⁇ 10 mm. It consists of 25 evenly spaced specimen plugs, each connected to a silicone catheter segment (Allegiance Healthcare Corp., McGaw Park, Ill.) whose anterior surface (0.3 cm 2 ) comes in contact with the flushed infusate.
- silicone catheter segment Allegiance Healthcare Corp., McGaw Park, Ill.
- the entire apparatus was gas sterilized using ethylene oxide.
- a 500 ml 5% dextrose in water (5% D 5 /W) was connected to the modified Robbins device through an intravenous tubing administration set and was subsequently infected with an innoculum of 10 8 CFU/ml of methicillin-resistant Staphylococcus aureus (MRSA), to produce an infected infusate at the concentration of 2 ⁇ 10 5 CFU/ml.
- MRSA methicillin-resistant Staphylococcus aureus
- the biofilm-producing S. aureus isolates were obtained from patients with CRBSI.
- 500 ml 5% D 5 /W bag was infected with a biofilm-producing C.
- the infected bag was removed and a 250 ml saline sterile bag which was infused through the MRD at 125 ml/hour for 2 hours in order to remove all free floating organisms.
- a 250 ml saline sterile bag which was infused through the MRD at 125 ml/hour for 2 hours in order to remove all free floating organisms.
- at least three catheter segments were randomly removed from the 25 evenly spaced catheter segments in the MRD and studied by scanning electron microscopy. This was repeated for every organism tested.
- the surface of the catheter segment that was exposed to the infected infusate was scraped with a sterile wooden applicator stick and placed, along with the stick, in a tube containing 0.5 ml of trypticase soy broth.
- the tubes were sonicated for five minutes; 0.1 ml of the sonicated broth solution in the tube was pipetted and plated over a blood agar plate, which was incubated at 37° for 24 hours.
- the agar plates were checked for any contaminants.
- the isolated organisms had to be of the same species and colonial morphology as the original organism used to infect the infusate.
- the number of colonies quantitated from the agar plate was multiplied by five to correct for the dilution factor and to determine the total number of colonies isolated from a particular catheter segment. A confluent growth of 100 or greater was calculated as ⁇ 500 colonies.
- Inhibitory activity or suppression is defined as no growth of microbial organisms immediately after 15 minute exposure to the antimicrobial solution. However, regrowth of the organisms after 24 hours incubation in broth was observed.
- Eradication is defined as no growth of organisms after immediate 15 minutes exposure to the antimicrobial solution with no subsequent growth upon reincubation for 24 hours in broth.
- Minocycline alone, EDTA alone or the combination of minocycline and EDTA failed to eradicate organisms embedded in biofilm after a rapid exposure of only 15 minutes.
- 25% ethanol solution also failed to eradicate organisms embedded in biofilm and a high level of regrowth was apparent after catheter segments were re-incubated in broth for an additional 24 hours at 37° C.
- the bacteria was added to form the biofilm.
- the plasma was suctioned out from the tubes and replaced with 0.5 ml of bacterial inoculum (50 ml of Mueller-Hinton broth containing 4-5 colonies of freshly grown bacteria). The tubes were incubated overnight at 37° C.
- a drug was added in an attempt to kill the bacteria.
- the pieces were washed in 0.5 ml of 0.9% saline in order to remove any planktonic bacteria.
- the tubes (containing the biofilm disks and saline) were placed in the incubator at 37° C. for 30 minutes.
- the saline was then pipetted out using sterile plastic transfer pipettes (taking care not to disturb the pieces too much).
- the silicone disks were then transferred to new 5 ml snap-top falcon tubes containing 0.5 ml of the drug solution to be tested.
- the drug solutions tested were as follows: (1) minocycline 3 mg/ml; (2) EDTA 30 mg/ml; (3) 25% ethanol solution; (4) EDTA 30 mg/ml in 25% ethanol; (5) minocycline 3 mg/ml in 25% ethanol; (6) minocycline 3 mg/ml with EDTA 30 mg/ml; and (7) triple combination of minocycline 3 mg/ml and EDTA 30 mg/ml in 25% ethanol solution.
- the disks were allowed to sit in the drug for 1 hour. The drug was then suctioned out using a plastic transfer pipette. The pieces were once again washed with 0.5 ml saline (added, and shaken for 30 seconds).
- the disks were then transferred to 15 ml snap-top falcon tubes containing 5 ml of 0.9% saline. The pieces were sonicated for 5 minutes, and then vortexed for 30 seconds. 100 microliters ( ⁇ l) of the saline was then plated on a room temperature TSAII blood agar plate, and evenly spread using a sterile glass spreader. The plates were incubated overnight at 37° C.
- the pieces were once again washed with 0.5 ml saline (added, and shaken for 30 seconds). The pieces were then transferred to new sterile 5 ml snap-top falcon tubes containing 0.5 ml of sterile trypticase soy broth (TSB) and then placed in the incubator at 37° C. overnight.
- TTB trypticase soy broth
- the silicone disk bioprosthetic colonization model has been previously described by Kuhn et al. (2002).
- This in vitro model is more clinically relevant than the modified Robin device, in vitro model, in that it allows the silicone disk segments to be immersed in serum prior to exposing to high inoculum of bacteria or fungi. Furthermore, it allows a higher concentration of adherence of bacteria and fungi on the silicone disk of up to 5,000 CFU/disk (the modified Robbins device allows for only 500 CFU/latex catheter segment). Because of the high inoculum that the silicone disk segments were exposed to in the bioprosthetic colonization model, the various disk segments were exposed to the various antimicrobial agents for one hour (rather than 15 minutes in the modified Robbins device).
- Table 5 shows a similar trend for Candida parapsilosis .
- Minocycline alone, EDTA alone, or the combination of M-EDTA failed to suppress or eradicate the growth of Candida parapsilosis on silicone disks. Furthermore, there was a heavy regrowth of the C. parapsilosis on silicone disks after exposure to these agents and reincubation for 24 hours. Twenty-five percent ethanol alone, EDTA in 25% ethanol or minocycline in 25% ethanol failed to completely suppress Candida parapsilosis growth after one hour exposure and there was heavy regrowth after 24 hour reincubation. The triple combination of M-EDTA in 25% ethanol completely eradicated the organisms on the silicone disks after 1 hour exposure. Furthermore, the level of regrowth associated with a triple combination after 24 hours of reincubation was significantly lower than all the other alternative agents or their dual combination.
- the two in vitro models of colonization show that the triple combination is uniquely and highly effective in eradicating organisms embedded in biofilm on latex and silicone polymers with minimal or no regrowth after 24 hour exposure to the combination.
- These two models are predictive of the clinical efficacy of this triple combination in eradicating organisms embedded in biofilm on catheters at a temperature of 37° C.
- the triple combination is superior in efficacy to the combination of minocycline and EDTA, EDTA and ethanol or minocycline and ethanol.
- compositions and/or methods and/or apparati disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and/or apparati and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.
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US14/253,265 Expired - Lifetime US9078441B2 (en) | 2003-06-06 | 2014-04-15 | Antimicrobial flush solutions |
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EP1644024A2 (en) | 2006-04-12 |
CY1122206T1 (el) | 2020-11-25 |
US20050013836A1 (en) | 2005-01-20 |
EP1644024B1 (en) | 2019-07-31 |
WO2004108091A3 (en) | 2005-05-12 |
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US20100055086A1 (en) | 2010-03-04 |
EP1644024A4 (en) | 2012-04-04 |
US20130164363A1 (en) | 2013-06-27 |
CA2528522A1 (en) | 2004-12-16 |
PT1644024T (pt) | 2019-10-24 |
US9078441B2 (en) | 2015-07-14 |
US20140228327A1 (en) | 2014-08-14 |
US8709342B2 (en) | 2014-04-29 |
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PL1644024T3 (pl) | 2020-03-31 |
US20110201692A1 (en) | 2011-08-18 |
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